Compositions for Enhancing Bioavailability of Pharmaceuticals, Supplements and Ingested Substances

ABSTRACT

The present invention relates to compositions and methods for enhancing bioavailability of health-promoting substances, such as pharmaceuticals and nutritional supplements. The subject invention utilizes an adjuvant composition comprising one or more microbial-produced biosurfactants and/or isoforms thereof, to enhance bioavailability of health-promoting substances and to reduce the effective dosage that is required.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationsNo. 62/537,502, filed Jul. 27, 2017, No. 62/647,974, Mar. 26, 2018, andNo. 62/690,365, filed Jul. 27, 2018, both of which are incorporated byreference herein in their entirety.

BACKGROUND OF THE INVENTION

In general, bioavailability can refer to the rate and extent to which asubstance reaches and enters a desired body system of a living organism,and can be effective therein. Specifically, bioavailability in thecontext of pharmacology is a measure of the rate and extent to which adrug reaches a site of action. In the realm of nutrition,bioavailability for food and dietary supplements can be defined as theproportion of an administered substance (or ingested substance) capableof being absorbed by the body and which is then available for use orstorage in the body. Furthermore, bioavailability can also be themeasure by which certain substances from the environment enter a livingorganism.

The bioavailability of a substance can play an important role in itsusefulness for a living organism, and can change based on a variety offactors. For example, the bioavailability of ingested substances can beaffected by the solubility of the substance, the rejection of thesubstance by the epithelium, or the speed at which the substance entersthrough the layers of the gastrointestinal (GI) tract. Substances withlow solubility may not have a sufficient retention time, as they areincapable of penetrating either through the cells or the tight junctionsbetween the cells of the GI tract. Thus, most, if not all of thesubstance is released from the body, unabsorbed and unused.

In addition to solubility, rejection of the substance is another factoraffecting bioavailability. For example, many substances can be rejectedby P-glycoprotein 1, a protein of the cell membrane that pumps foreignsubstances out of cells. More formally, it is an ATP-dependent effluxpump with broad substrate specificity. This pump is thought to haveevolved as a defense mechanism against harmful substances, but can serveas an obstacle in many cases when a foreign, yet desirable, substance issought to be introduced into the body. It is broadly distributed andexpressed in the cells of a variety of organs, including the intestinalepithelium, where it pumps, for example, xenobiotics, back into theintestinal lumen; in liver cells, where it pumps substances into bileducts; in the cells of the proximal tubule of the kidney, where it pumpssubstances into the urine-conducting ducts; and in the capillaryendothelial cells composing the blood-brain barrier and blood-testisbarrier, where it pumps substances back into the capillaries.

Pharmaceuticals, supplements and nutrition are important aspects ofleading a healthy life; however, the dosage or amount of certainhealth-promoting substances that must be administered to a subject isoften far greater than is actually needed to have a desired effect. Thisis because evolutionary obstacles hinder the bioavailability of certaincompounds and nutrients from reaching a desired site of action, forexample, through epithelial cells and through the blood-brain barrier.

Thus, there is a need for compositions and methods that are capable ofenhancing the bioavailability of a broad range of pharmaceuticals,supplements, nutrients and other health-promoting substances that areadministered to a subject.

BRIEF SUMMARY OF THE INVENTION

The subject invention provides materials and methods for improving thebioavailability of pharmaceuticals, supplements, nutrients and/or otherhealth-promoting substances. In particular, the subject inventionprovides adjuvant compositions comprising microbial growth by-productsfor use in enhancing bioavailability of health-promoting substances.Advantageously, the microbe-based products and methods of the subjectinvention are non-toxic and cost-effective.

In certain specific embodiments, the subject invention providesapproaches to enhancing bioavailability of a health-promoting substanceusing microbial growth by-products by, for example, suppressingP-glycoproteins and/or modulating other physical barrier mechanisms thatwould otherwise reduce the penetration of certain substances into, forexample, a subject's epithelial cells and/or across the blood-brainbarrier (BBB).

In one embodiment, adjuvant compositions are provided for enhancingbioavailability of a health-promoting compound, wherein said adjuvantcompositions comprise therapeutically-effective amounts of one or morebiosurfactants.

Biosurfactants are surface-active substances produced by microorganismsthat lower the surface tension (or interfacial tension) between twoliquids or between a liquid and a solid. All biosurfactants areamphiphiles. They consist of two parts: a polar (hydrophilic) moiety andnon-polar (hydrophobic) group. Due to their amphiphilic structure,biosurfactants increase the surface area of hydrophobic water-insolublesubstances, increase the water bioavailability of such substances, andchange the properties of bacterial cell surfaces. Furthermore,biosurfactants accumulate at interfaces, and reduce the surface andinterfacial tension between the molecules of liquids, solids, and gases,thus leading to the formation of aggregated micellular structures insolution.

Biosurfactants according to the subject invention include low molecularweight glycolipids (e.g., sophorolipids, rhamnolipids,mannosylerythritol lipids and trehalose lipids), lipopeptides (e.g.,surfactin, iturin, fengycin and lichenysin), flavolipids, phospholipids,and high molecular weight polymers such as lipoproteins,lipopolysaccharide-protein complexes, and polysaccharide-protein-fattyacid complexes. In certain embodiments, the one or more biosurfactantsare isolated and/or purified.

The one or more biosurfactants can further include any one or acombination of: a modified form, derivative, fraction, isoform orsubtype of a biosurfactant, including forms that are naturally orartificially modified. The use of different isomers or forms of thebiosurfactants is beneficial in that the skilled artisan can tailor theadjuvant composition depending upon its interactions with a particularhealth-promoting compound. That is, certain isoforms of a biosurfactantmight be more effective with certain health-promoting compounds due to,for example, the chemical structure of the compound.

Preferably, the one or more biosurfactants are present in the subjectadjuvant composition in critical micelle concentration (CMC).

In some embodiments, the adjuvant composition further comprises atherapeutically-effective dose of a target health-promoting compound,the bioavailability of which is sought to be enhanced. In thisembodiment, the adjuvant composition can be mixed with thehealth-promoting compound. Alternatively, the adjuvant composition canbe a separate composition from the target health-promoting compound,wherein the adjuvant composition is intended to be administered to asubject separately, but close in time to, the health-promoting compound.

The health-promoting compound can be, for example, a pharmaceuticalcompound, a nutritional supplement, or even simply water. In oneembodiment, the subject compositions are formulated as anorally-consumable product, such as, for example, a capsule, a pill or adrinkable liquid. In another embodiment, the subject compositions areformulated to be administered via injection, suppository, inhalation, orany other mode of administration.

In one embodiment, the subject invention provides a delivery system fora health-promoting compound, wherein the biosurfactants of the adjuvantcomposition form a liposome or nanocapsule with the health-promotingcompound encapsulated therein. In one embodiment, additional biologicalpolymers can be included to provide further structure for thenanocapsule.

This nanocapsule delivery system can enhance the bioavailability of ahealth-promoting compound by protecting the compound from components inthe blood, such as proteins and other molecules, that otherwise mightbind to and/or degrade the compound and prevent it from arriving at atarget site. The nanocapsule delivery system can allow forhealth-promoting compounds that might otherwise by degraded by acids orenzymes in the GI tract to be administered orally, as it creates abarrier against the acids or enzymes. In this regard, the deliverysystem can comprise an enteric coating. Furthermore, the nanocapsuledelivery system formulation allows for time release of thehealth-promoting compound, thereby reducing the potential toxicity orpotential negative side-effects of a compound in a subject.

The subject invention further provides a method of enhancing thebioavailability of a health-promoting compound, which comprisesadministering a therapeutically-effective amount of an adjuvantcomposition of the subject invention to a subject in need thereof andadministering a therapeutically-effective amount of the health-promotingcompound to the subject.

The methods can further be used to allow for oral administration ofhealth-promoting compounds that might otherwise by degraded by acids orenzymes in the GI tract. Furthermore, the methods can be used to reducethe dosage required for a health-promoting compound, and reduce thepotential toxicity or potential negative side-effects of a compound in asubject.

In preferred embodiments, the adjuvant composition comprises one or morebiosurfactants, including any modified form, derivative, fraction,isoform or subtype of biosurfactants selected from, for example, asophorolipid, rhamnolipid, mannosylerythritol lipid, trehalose lipid,surfactin, iturin, fengycin and lichenysin. The health-promotingcompound can be administered simultaneously with the adjuvantcomposition or otherwise close in time to administering the adjuvantcomposition.

Health-promoting compounds (or health-promoting substances) comprise anymolecule or molecules that are meant to be delivered into blood and/orlymphatic circulation, as well as into tissues and organs, andultimately reach a site in a subject's body where a positive impact onthe subject's health can be effected. Non-limiting examples ofhealth-promoting compounds include pharmaceuticals and/or nutritionalsupplements categorized as pain-relievers, antihistamines, antivirals,anticancer and/or chemotherapeutic compounds, antibiotics,antimicrobials, antiseizure compounds, anti-inflammatory compounds,vaccines, cholesterol-lowering compounds, antidepressants, vitamins,minerals, nutrients, water and many others.

In one embodiment, the health-promoting substance is an orallydeliverable health-promoting substance, which, in particular, is anymolecule or molecules that is delivered via initial absorption into thegastrointestinal tract or into the mucus membranes of the mouth (e.g.,by way of sublingual or buccal administration).

Advantageously, the materials and methods of the subject invention canhelp improve the quality of life for individuals who are eithersuffering from a particular health condition or who are already healthy(e.g., generally free from illness or injury) but are simply seeking toenhance their state of being. Furthermore, the subject invention can beused to reduce the dosage of certain pharmaceuticals and/or supplementsthat are required to be considered therapeutically-effective, thusreducing the cost and potential toxicity and/or negative side-effectsthat might arise from administering them to a subject.

DETAILED DESCRIPTION

The subject invention provides materials and methods for improving thebioavailability of pharmaceuticals, supplements, nutrients and/or otherhealth-promoting substances. In particular, the subject inventionprovides adjuvant compositions comprising microbial growth by-productsfor use in enhancing bioavailability of health-promoting substances.Advantageously, the microbe-based products and methods of the subjectinvention are non-toxic and cost-effective.

Further described herein are approaches to enhancing bioavailability ofa health-promoting compound, which utilize microbial growth by-productsto, for example, suppress P-glycoproteins and modulate other bloodplasma proteins and/or physical barrier mechanisms that reduce thepenetration of certain compounds into a subject's epithelial cellsand/or across the BBB. The subject invention also provides methods forreducing the dosage of a health-promoting compound required for thehealth-promoting compound to be therapeutically-effective in a subject.

Selected Definitions

As used herein, the term “adjuvant” in the context of the subjectcompositions means an auxiliary compound that can aid in, contribute to,and/or enhance the effectiveness of a substance that is administeredwith the adjuvant. For example, an adjuvant can be taken alongside orincluded in a prescription drug or a supplement to aid in theeffectiveness of the active, primary active ingredient(s), whatever thepurpose may be (e.g., treating a disease or enhancing the functioning ofan organ or system in the body).

As used herein, the term “subject” refers to an animal, such as amammal, needing or desiring delivery of the benefits provided by ahealth-promoting substance. The animal may be for example, pigs, horses,goats, cats, mice, rats, dogs, apes, fish, chimpanzees, orangutans,guinea pigs, hamsters, cows, sheep, birds, e.g., chickens, as well asany other vertebrate or invertebrate. These benefits can include, butare not limited to, treatment of a health condition, disease ordisorder; prevention of a health condition, disease or disorder;hydration or rehydration; nutritional enhancement and/or supplementationfor, e.g., athletic performance or weight control; immune health;enhancement of function of an organ, tissue or system in the body;and/or simply pleasure. The preferred subject in the context of thisinvention is a human, either male or female. In some embodiments, asubject is suffering from a health condition, disease or disorder, whilein some embodiments, the subject is in a state of good health (i.e.,free from injury or illness), but desires enhanced health and/orfunctioning of an particular organ, tissue or body system. The subjectcan be of any age or stage of development, including infant, toddler,adolescent, teenager, adult, and senior.

As used herein, the terms “therapeutically-effective amount,”“therapeutically-effective dose,” “effective amount,” and “effectivedose” are used to refer to an amount or dose of a compound orcomposition that, when administered to a subject, is capable of treatingor improving a condition, disease or disorder in a subject, or that iscapable of providing enhancement in health or function to an organ,tissue or body system. In other words, when administered to a subject,the amount is “therapeutically effective.” The actual amount will varydepending on a number of factors including, but not limited to, theparticular condition, disease or disorder being treated or improved; theseverity of the condition; the particular organ, tissue or body systemof which enhancement in health or function is desired; the size, age,and health of the patient; and the route of administration.

As used herein, the term “treatment” refers to eradicating, reducing,ameliorating, or reversing a sign or symptom of a health condition,disease or disorder to any extent, and includes, but does not require, acomplete cure of the condition, disease or disorder. Treating can becuring, improving, or partially ameliorating a disorder. “Treatment” canalso include improving or enhancing a condition or characteristic, forexample, bringing the function of a particular system in the body to aheightened state of health or homeostasis.

As used herein, “preventing” a health condition, disease or disorderrefers to avoiding, delaying, forestalling, or minimizing the onset of aparticular sign or symptom of the condition, disease or disorder.Prevention can, but is not required to be, absolute or complete, meaningthe sign or symptom may still develop at a later time. Prevention caninclude reducing the severity of the onset of such a condition, diseaseor disorder, and/or inhibiting the progression of the condition, diseaseor disorder to a more severe condition or disorder.

As used herein, reference to a “microbe-based composition” means acomposition that comprises components that were produced as the resultof the growth of microorganisms or other cell cultures. A microbe-basedcomposition may comprise the microbes themselves, or the microbes may beseparated from the broth in which they were cultivated, and thecomposition comprises residual cellular components and/or by-products ofmicrobial growth. Preferably, the compositions according to the subjectinvention have been separated from the microbes. The by-products ofmicrobial growth may be, for example, metabolites (e.g.,biosurfactants), cell membrane components, expressed proteins, and/orother cellular components.

The subject invention further provides “microbe-based products,” whichare products that are to be applied in practice to achieve a desiredresult. The microbe-based product can be simply the microbe-basedcomposition harvested from the microbe cultivation process.Alternatively, the microbe-based product may comprise furtheringredients that have been added. These additional ingredients caninclude, for example, stabilizers, buffers and/or appropriate carriers(e.g., water or salt solutions). The microbe-based product may comprisemixtures of microbe-based compositions. The microbe-based product mayalso comprise one or more components of a microbe-based composition thathave been processed in some way such as, but not limited to, filtering,centrifugation, lysing, drying, purification and the like.

As used herein, “harvested” refers to removing some or all of themicrobe-based composition from a growth vessel.

As used herein, an “isolated” or “purified” nucleic acid molecule,polynucleotide, polypeptide, protein or organic compound such as a smallmolecule (e.g., those described below), is substantially free of othercompounds, such as cellular material, with which it is associated innature. In certain embodiments, purified compounds are at least 60% byweight (dry weight) the compound of interest. Preferably, thepreparation is at least 75%, more preferably at least 90%, and mostpreferably at least 99%, by weight the compound of interest. Forexample, a purified compound is one that is at least 90%, 91%, 92%, 93%,94%, 95%, 98%, 99%, or 100% (w/w) of the desired compound by weight.Purity is measured by any appropriate standard method, for example, bycolumn chromatography, thin layer chromatography, or high-performanceliquid chromatography (HPLC) analysis. A purified or isolatedpolynucleotide (ribonucleic acid (RNA) or deoxyribonucleic acid (DNA))is free of the genes or sequences that flank it in itsnaturally-occurring state. A purified or isolated polypeptide is free ofthe amino acids or sequences that flank it in its naturally-occurringstate.

A “metabolite” refers to any substance produced by metabolism (i.e., agrowth by-product) or a substance necessary for taking part in aparticular metabolic process. A metabolite can be an organic compoundthat is a starting material (e.g., glucose), an intermediate (e.g.,acetyl-CoA) in, or an end product (e.g., n-butanol) of metabolism.Examples of metabolites include, but are not limited to, biosurfactants,enzymes, acids, solvents, gasses, alcohols, proteins, vitamins,minerals, microelements, amino acids, and polymers.

By “modulate” is meant alter (increase or decrease). Such alterationsare detected by standard art known methods such as those describedherein.

Ranges provided herein are understood to be shorthand for all of thevalues within the range. For example, a range of 1 to 20 is understoodto include any number, combination of numbers, or sub-range from thegroup consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, or 20 as well as all intervening decimal values between theaforementioned integers such as, for example, 1.1, 1.2, 1.3, 1.4, 1.5,1.6, 1.7, 1.8, and 1.9. With respect to sub-ranges, “nested sub-ranges”that extend from either end point of the range are specificallycontemplated. For example, a nested sub-range of an exemplary range of 1to 50 may comprise 1 to 10, 1 to 20, 1 to 30, and 1 to 40 in onedirection, or 50 to 40, 50 to 30, 50 to 20, and 50 to 10 in the otherdirection.

By “reduces” is meant a negative alteration of at least 1%, 5%, 10%,25%, 50%, 75%, or 100%.

By “reference” is meant a standard or control condition.

As used herein, a “biofilm” is a complex aggregate of microorganisms,such as bacteria, wherein the cells adhere to each other. The cells inbiofilms are physiologically distinct from planktonic cells of the sameorganism, which are single cells that can float or swim in liquidmedium.

As used herein, “surfactant” refers to a surface-active substance thatlowers the surface tension (or interfacial tension) between two liquidsor between a liquid and a solid. Surfactants act as, for example,detergents, wetting agents, emulsifiers, foaming agents, and/ordispersants. A surface-active substance produced by microorganisms isreferred to as a “biosurfactant.”

The transitional term “comprising,” which is synonymous with“including,” or “containing,” is inclusive or open-ended and does notexclude additional, unrecited elements or method steps. By contrast, thetransitional phrase “consisting of” excludes any element, step, oringredient not specified in the claim. The transitional phrase“consisting essentially of” limits the scope of a claim to the specifiedmaterials or steps “and those that do not materially affect the basicand novel characteristic(s)” of the claimed invention, e.g., the abilityto improve the bioavailability of a substance.

Unless specifically stated or obvious from context, as used herein, theterm “or” is understood to be inclusive. Unless specifically stated orobvious from context, as used herein, the terms “a,” “an” and “the” areunderstood to be singular or plural.

Unless specifically stated or obvious from context, as used herein, theterm “about” is understood as within a range of normal tolerance in theart, for example within 2 standard deviations of the mean. About can beunderstood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%,0.1%, 0.05%, or 0.01% of the stated value.

The recitation of a listing of chemical groups in any definition of avariable herein includes definitions of that variable as any singlegroup or combination of listed groups. The recitation of an embodimentfor a variable or aspect herein includes that embodiment as any singleembodiment or in combination with any other embodiments or portionsthereof.

Any compositions or methods provided herein can be combined with one ormore of any of the other compositions and methods provided herein.

Other features and advantages of the invention will be apparent from thefollowing description of the preferred embodiments thereof, and from theclaims. All references cited herein are hereby incorporated byreference.

Formulation and Delivery of Adjuvant Composition

The subject invention provides adjuvant compositions comprisingmicrobial growth by-products for use in enhancing bioavailability ofpharmaceuticals, supplements, nutrients and other health-promotingsubstances. Advantageously, the microbe-based products and methods ofthe subject invention are non-toxic and cost-effective.

In one embodiment, adjuvant compositions are provided for enhancingbioavailability of a health-promoting compound, wherein said adjuvantcompositions comprise therapeutically-effective amounts of one or morebiosurfactants.

In some embodiments, the adjuvant composition further comprises atherapeutically-effective dose of a health-promoting compound, thebioavailability of which is sought to be enhanced. In this embodiment,the adjuvant and the health-promoting compound are included together asone formulation with other optional additives.

Alternatively, the adjuvant composition can be administered to a subjectseparately from, but close in time to (e.g., five minutes or less beforeor after), administration of a target health-promoting compound.

Advantageously, the adjuvant composition of the subject invention iscapable of reducing a liquid's surface tension and reducing theinterfacial tension between liquid-liquid and liquid-solid interfaces.Additionally, a target health-promoting compound can exhibit resistanceto degradation by digestive juices (e.g., acids and enzymes) whenadministered into the gastrointestinal (GI) system along with thesubject adjuvant composition. Furthermore, in certain embodiments, thesubject adjuvant composition can help suppress and/or modulate theactivity of, for example, blood plasma proteins, P-glycoproteins, andother barriers and cell junctions that prevent certain compounds frompenetrating into a target site of the body.

The adjuvant composition preferably comprises biosurfactants, which aresurface-active substances produced by microorganisms. The biosurfactantsuseful according to the subject invention are safe, biodegradable andcan be produced with ease at low cost using selected organisms in or onrenewable substrates.

All biosurfactants are amphiphiles. They consist of two parts: a polar(hydrophilic) moiety and non-polar (hydrophobic) group. Due to theiramphiphilic structure, biosurfactants increase the surface area ofhydrophobic water-insoluble substances, increase the waterbioavailability of such substances, and change the properties ofbacterial cell surfaces. Furthermore, biosurfactants accumulate atinterfaces, and reduce the surface and interfacial tension between themolecules of liquids, solids, and gases, thus leading to the formationof aggregated micellular structures in solution.

Most biosurfactant-producing organisms produce biosurfactants inresponse to the presence of a hydrocarbon source (e.g., oils, sugar,glycerol, etc.) in the growing media. Other media components such asconcentration of iron can also affect biosurfactant productionsignificantly. Microbial biosurfactants are produced by a variety ofmicroorganisms such as bacteria, fungi, and yeasts, such as, forexample, Pseudomonas spp. (P. aeruginosa, P. putida, P. florescens, P.fragi, P. syringae); Flavobacterium spp.; Bacillus spp. (B. subtilis, B.pumillus, B. cereus, B. licheniformis); Wickerhamomyces spp. (Wanomalus), Candida spp. (C. albicans, C. rugosa, C. tropicalis, C.lipolytica, C. torulopsis); Rhodococcus spp.; Arthrobacter spp.;campylobacter spp.; cornybacterium spp.; Pichia spp. (P. anomala, P.occidentalis); Starmerella spp. (S. bombicola); and so on.

The biosurfactants may be obtained by fermentation processes known inthe art, e.g., solid-state fermentation, submerged fermentation, orcombinations thereof. The biosurfactant produced by microorganisms ofinterest may be retained in the microorganisms or secreted into theirgrowth medium. The growth medium may contain compounds that stabilizethe activity of the biosurfactant. Furthermore, the growth by-productmay be isolated, concentrated and/or purified.

Biosurfactants according to the subject invention include low molecularweight glycolipids (e.g., sophorolipids, rhamnolipids,mannosylerythritol lipids and trehalose lipids), lipopeptides (e.g.,surfactin, iturin, fengycin and lichenysin), flavolipids, phospholipids,and high molecular weight polymers such as lipoproteins,lipopolysaccharide-protein complexes, and polysaccharide-protein-fattyacid complexes.

The one or more biosurfactants can further include one or a combinationof: a modified form, derivative, fraction, isoform or subtype of abiosurfactant, including forms that are naturally or artificiallymodified. The use of different isomers or forms of biosurfactants isbeneficial in that the skilled artisan can tailor the adjuvantcomposition depending upon its interactions with a particularhealth-promoting compound. That is, certain isoforms of a biosurfactantmight be more effective with certain health-promoting compounds due to,for example, the chemical structure of the compound.

In certain embodiments, the biosurfactant is a sophorolipid (SLP), suchas, for example, a lactonic or acidic form sophorolipid, anon-acetylated sophorolipid, a mono-acetylated sophorolipid, adi-acetylated sophorolipid, or any other isoform thereof.

In certain embodiments, the biosurfactant is a rhamnolipid (RLP), suchas, for example, a mono-rhamnolipid, a di-rhamnolipid, or any otherisoform thereof.

In certain embodiments, the biosurfactant is a mannosylerythritol lipid(MEL), such as, for example, MEL-A, MEL-B, MEL-C, or MEL-D, or any otherisoforms with varying fatty acid lengths and/or hydrophobic portions.

In certain embodiments, the biosurfactant is a trehalose lipid (TL) orany other isoform thereof.

In certain embodiments, the biosurfactant is a lipopeptide, includinglinear or cyclic form lipopeptides, or any other isoforms thereof. As anexample, surfactin is a lipopeptide that can have a structure comprisinga peptide loop of seven amino acids and a hydrophobic fatty acid chainthirteen to fifteen carbons long. In an exemplary embodiment, the aminoacids comprise L-aspartic acid, L-leucine, glutamic acid, L-leucine,L-valine and two D-leucines.

As another example, iturin is a lipopeptide with a structure comprisinga peptide loop of seven amino acids and a β-amino fatty acid chain thatcan vary from 14 to 17 carbons long. In one embodiment, iturin A isutilized according to the subject invention.

The biosurfactants are preferably present in the adjuvant composition intherapeutically-effective amounts. In one embodiment, this means thebiosurfactants are present in critical micelle concentration (CMC). CMCis the concentration of surfactants above which micelles will form andall additional surfactants added to the system either convert tomicelles or add to the existing micelles.

In certain embodiments, a therapeutically-effective amount ofbiosurfactants in the composition is 0.001 to 90% by weight (wt %),preferably 50 wt % or less, more preferably 25 wt % or less, even morepreferably 10 wt % or less. In certain embodiments, the biosurfactant ispresent at more than 0.01, 0.02, 0.03, 0.05, 0.08, 0.1, 0.2, or 0.5%.

In some embodiments, the adjuvant composition further comprises atherapeutically-effective dose of a target health-promoting compound,the bioavailability of which is sought to be enhanced. In thisembodiment, the adjuvant composition can be mixed with thehealth-promoting compound. Alternatively, the adjuvant composition canbe a separate composition from the target health-promoting compound,wherein the adjuvant composition is intended to be administered to asubject at the same time as the health-promoting compound.

The health-promoting compound can be, for example, a pharmaceuticalcompound, a nutritional supplement, or even simply water. In oneembodiment, the subject compositions are formulated as anorally-consumable product, such as, for example, a capsule, a pill or adrinkable liquid. In another embodiment, the subject compositions areformulated to be administered via injection, inhalation, or any othermode of administration.

The subject invention is useful for enhancing the bioavailability of“health-promoting compounds” or “health-promoting substances,” whichcomprise any molecule or molecules that are meant to be delivered intoblood and/or lymphatic circulation, as well as into tissues and organs,and ultimately reach a site in a subject's body where a positive impacton the subject's health, either locally or systemically, can beeffected. Health-promoting compounds include, for example, any categoryof supplement and/or pharmaceutical (including biopharmaceuticals) usedfor, for example, relieving pain, fever and/or inflammation; reducingthe symptoms of allergies or colds; suppressing or treating a virus;treating cancer; treating a microbial infection; suppressing orpreventing seizures; lowering or managing cholesterol; managingdiabetes; treating depression or anxiety; hydrating or rehydrating;controlling body weight; enhancing athletic performance; and reducing orenhancing fertility, to name just a few.

In one embodiment, the adjuvant composition is formulated as a deliverysystem for a health-promoting compound, wherein the biosurfactants ofthe adjuvant composition form a liposome or nanocapsule with thehealth-promoting compound encapsulated therein. In one embodiment,additional biological polymers can be included to provide furtherstructure for the nanocapsule.

This nanocapsule delivery system can enhance the bioavailability of ahealth-promoting compound by protecting the compound from components inthe blood, such as proteins and other molecules, that otherwise mightbind to the compound and prevent it from penetrating a target site.Additionally, the nanocapsule delivery system can allow forhealth-promoting compounds that might otherwise by degraded by acids orenzymes in the GI tract to be administered orally, as it creates abarrier against the acids or enzymes. Furthermore, the nanocapsuledelivery system formulation allows for time release of thehealth-promoting compound, thereby reducing the potential toxicity orpotential negative side-effects of a compound in a subject.

In one embodiment, the adjuvant composition can be formulated tocomprise an orally deliverable health-promoting substance and/or to beadministered simultaneously with one as an orally consumable product. Anorally deliverable health-promoting substance is any physiologicallyactive substance delivered via initial absorption into thegastrointestinal tract, or into the mucus membranes of the mouth (e.g.,by way of sublingual or buccal administration).

Orally consumable products according to the invention are anypreparations or compositions suitable for consumption, for nutrition,for oral hygiene or for pleasure, and are products intended to beintroduced into the human or animal oral cavity, to remain there for acertain period of time and then to either be swallowed (e.g., food readyfor consumption or pills) or to be removed from the oral cavity again(e.g., chewing gums or products of oral hygiene or medical mouthwashes). While an orally-deliverable health-promoting substance can beformulated into an orally consumable product, and an orally consumableproduct can comprise an orally-deliverable health-promoting substance,the two terms are not meant to be used interchangeably herein.

Orally consumable products include all substances or products intendedto be ingested by humans or animals in a processed, semi-processed orunprocessed state. This also includes substances that are added toorally-consumable products (particularly food and pharmaceuticalproducts) during their production, treatment or processing and intendedto be introduced into the human or animal oral cavity.

Orally-consumable products can also include substances intended to beswallowed by humans or animals and then digested in an unmodified,prepared or processed state; the orally consumable products according tothe invention therefore also include casings, coatings or otherencapsulations that are intended also to be swallowed together with theproduct or for which swallowing is to be anticipated.

In one embodiment, the orally-consumable product is a capsule, pill,syrup, emulsion or liquid suspension containing a desiredorally-deliverable substance. In one embodiment, the orally-consumableproduct can comprise an orally-deliverable substance in powder form,which can be mixed with water or another liquid to produce a drinkableorally-consumable product.

In some embodiments, the orally-consumable product according to theinvention can comprise one or more formulations intended for nutritionor pleasure. These particularly include baking products (e.g., bread,dry biscuits, cake, and other pastries), sweets (e.g., chocolates,chocolate bar products, other bar products, fruit gum, coated tablets,hard caramels, toffees and caramels, and chewing gum), alcoholic ornon-alcoholic beverages (e.g., cocoa, coffee, green tea, black tea,black or green tea beverages enriched with extracts of green or blacktea, Rooibos tea, other herbal teas, fruit-containing lemonades,isotonic beverages, soft drinks, nectars, fruit and vegetable juices,and fruit or vegetable juice preparations), instant beverages (e.g.,instant cocoa beverages, instant tea beverages, and instant coffeebeverages), meat products (e.g., ham, fresh sausage preparations or rawsausage preparations, and seasoned oder, marinated fresh meat or saltedmeat products), eggs or egg products (e.g., dried whole egg, egg white,and egg yolk), cereal products (e.g., breakfast cereals, muesli bars,and pre-cooked instant rice products), dairy products (e.g., whole fator fat reduced or fat-free milk beverages, rice pudding, yoghurt, kefir,cream cheese, soft cheese, hard cheese, dried milk powder, whey, butter,buttermilk, and partly or wholly hydrolyzed products containing milkproteins), products from soy protein or other soy bean fractions (e.g.,soy milk and products prepared thereof, beverages containing isolated orenzymatically treated soy protein, soy flour containing beverages,preparations containing soy lecithin, fermented products such as tofu ortempeh products prepared thereof and mixtures with fruit preparationsand, optionally, flavoring substances), fruit preparations (e.g., jams,fruit ice cream, fruit sauces, and fruit fillings), vegetablepreparations (e.g., ketchup, sauces, dried vegetables, deep-freezevegetables, pre-cooked vegetables, and boiled vegetables), snackarticles (e.g., baked or fried potato chips (crisps) or potato doughproducts, and extrudates on the basis of maize or peanuts), products onthe basis of fat and oil or emulsions thereof (e.g., mayonnaise,remoulade, and dressings), other ready-made meals and soups (e.g., drysoups, instant soups, and pre-cooked soups), seasonings (e.g.,sprinkle-on seasonings), sweetener compositions (e.g., tablets, sachets,and other preparations for sweetening or whitening beverages or otherfood). The present compositions may also serve as semi-finished productsfor the production of other compositions intended for nutrition orpleasure.

In one embodiment, the adjuvant composition can be formulated tocomprise a health-promoting substance and/or to be administeredsimultaneously with one via a route of administration, including, forexample, injection (e.g., intravenous (IV), intramuscular (IM),intraperitoneal, intrathecal or subcutaneous), transdermal, rectal,urogenital (e.g., vaginal), ocular, aural, nasal, inhalation andcutaneous routes.

The subject composition can further comprise one or morepharmaceutically acceptable carriers and/or excipients, and can beformulated into preparations in, for example, solid, semi-solid, liquidor gaseous forms, such as tablets, capsules, powders, granules,ointments, gels, lotions, solutions, suppositories, drops, patches,injections, inhalants and aerosols.

The term “pharmaceutically acceptable” as used herein means compatiblewith the other ingredients of a pharmaceutical composition and notdeleterious to the recipient thereof.

Carriers and/or excipients according the subject invention can includeany and all solvents, diluents, buffers (such as, e.g., neutral bufferedsaline, phosphate buffered saline, or optionally Tris-HCl, acetate orphosphate buffers), oil-in-water or water-in-oil emulsions, aqueouscompositions with or without inclusion of organic co-solvents suitablefor, e.g., IV use, solubilisers (such as, e.g., Tween 80, Polysorbate80), colloids, dispersion media, vehicles, fillers, chelating agents(such as, e.g., EDTA or glutathione), amino acids (such as, e.g.,glycine), proteins, disintegrants, binders, lubricants, wetting agents,emulsifiers, sweeteners, colorants, flavorings, aromatisers, thickeners,coatings, preservatives (such as, e.g., Thimerosal, benzyl alcohol),antioxidants (such as, e.g., ascorbic acid, sodium metabisulfite),tonicity controlling agents, absorption delaying agents, adjuvants,bulking agents (such as, e.g., lactose, mannitol) and the like. The useof carriers and/or excipients in the field of drugs and supplements iswell known. Except for any conventional media or agent that isincompatible with the target health-promoting substance or with theadjuvant composition, its use in the subject compositions may becontemplated.

In one embodiment, the adjuvant composition can be made into aerosolformulations so that, for example, it can be nebulized or inhaled.Suitable pharmaceutical formulations for administration in the form ofaerosols or sprays are, for example, solutions, suspensions oremulsions. Formulations for oral or nasal aerosol or inhalationadministration may also be formulated with illustrative carriers,including, for example, saline, polyethylene glycol or glycols, DPPC,methylcellulose, or in mixture with powdered dispersing agents orfluorocarbons. Aerosol formulations can be placed into pressurizedpropellants, such as dichlorodifluoromethane, propane, nitrogen,fluorocarbons, and/or other solubilizing or dispersing agents known inthe art. Illustratively, delivery may be by use of a single-use deliverydevice, a mist nebulizer, a breath-activated powder inhaler, an aerosolmetered-dose inhaler (MDI) or any other of the numerous nebulizerdelivery devices available in the art. Additionally, mist tents ordirect administration through endotracheal tubes may also be used.

In one embodiment, the adjuvant composition can be formulated foradministration via injection, for example, as a solution or suspension.The solution or suspension can comprise suitable non-toxic,parenterally-acceptable diluents or solvents, such as mannitol,1,3-butanediol, water, Ringer's solution or isotonic sodium chloridesolution, or suitable dispersing or wetting and suspending agents, suchas sterile, bland, fixed oils, including synthetic mono- ordiglycerides, and fatty acids, including oleic acid. One illustrativeexample of a carrier for intravenous use includes a mixture of 10% USPethanol, 40% USP propylene glycol or polyethylene glycol 600 and thebalance USP Water for Injection (WFI). Other illustrative carriers forintravenous use include 10% USP ethanol and USP WFI; 0.01-0.1%triethanolamine in USP WFI; or 0.01-0.2% dipalmitoyldiphosphatidylcholine in USP WFI; and 1-10% squalene or parenteralvegetable oil-in-water emulsion. Water or saline solutions and aqueousdextrose and glycerol solutions may be preferably employed as carriers,particularly for injectable solutions. Illustrative examples of carriersfor subcutaneous or intramuscular use include phosphate buffered saline(PBS) solution, 5% dextrose in WFI and 0.01-0.1% triethanolamine in 5%dextrose or 0.9% sodium chloride in USP WFI, or a 1 to 2 or 1 to 4mixture of 10% USP ethanol, 40% propylene glycol and the balance anacceptable isotonic solution such as 5% dextrose or 0.9% sodiumchloride; or 0.01-0.2% dipalmitoyl diphosphatidylcholine in USP. WFI and1 to 10% squalene or parenteral vegetable oil-in-water emulsions.

Further components can be added to the compositions as are determined bythe skilled artisan such as, for example, buffers, carriers, viscositymodifiers, preservatives, flavorings, dyes and other ingredientsspecific for an intended use. One skilled in this art will recognizethat the above description is illustrative rather than exhaustive.Indeed, many additional formulations techniques andpharmaceutically-acceptable excipients and carrier solutions suitablefor particular modes of administration are well-known to those skilledin the art.

Methods of Enhancing Bioavailability of Health-Promoting Substances

The subject invention further provides methods of enhancing thebioavailability of a health-promoting substance in a subject in needthereof, which comprises administering a therapeutically-effectiveamount of an adjuvant composition comprising one or more biosurfactantsto the subject and administering a therapeutically-effective amount ofthe health-promoting compound to the subject.

The method can also be used to reduce the dosage required for ahealth-promoting compound, and reduce the potential toxicity orpotential negative side-effects of a compound in a subject. Furthermore,the method can be used to allow for oral administration ofhealth-promoting compounds that might otherwise by degraded by acids orenzymes in the GI tract

The health-promoting compound can be administered simultaneously withthe adjuvant composition, for example, as part of a single formulation.In one embodiment, the method comprises administering thehealth-promoting compound to the subject using the adjuvant nanocapsuledelivery system described herein.

Alternatively, the health-promoting compound can be administeredseparately from the adjuvant composition. In this alternate embodiment,the health-promoting compound is administered either immediately beforeor immediately after the adjuvant composition is administered, wherein“immediately before” or “immediately after” means. 5 minutes, 4 minutes,3 minutes, 2 minutes, 1 minute, 30 seconds or less before or after.

In preferred embodiments, the biosurfactants of the adjuvant compositionare selected from, for example, glycolipids (e.g., SLP, RLP, MEL andTL), lipopeptides (e.g., surfactin, iturin, fengycin and lichenysin),and any modified form, derivative, fraction, isoform or subtype thereof.Combinations of biosurfactants and their various forms are alsoenvisioned.

As used herein, “administering” a composition or product refers todelivering it to a subject such that it contacts a target or site suchthat the composition or product can have an effect on that target orsite. The effect can be due to, for example, the action of ahealth-promoting compound, or due to a biosurfactant composition.Administration can be acute or chronic (e.g., daily, weekly, monthly,etc.) or in combination with other agents. The subject adjuvantcomposition, whether administered in the same formulation as the targethealth-promoting compound or within, for example, 5 minutes of thetarget compound, can be administered by any route of administrationprovided it is formulated for such a route. In this way, the therapeuticeffects attainable by the methods and compositions of the invention canbe, for example, systemic, local, tissue-specific, etc., depending ofthe specific needs of a given application of the invention.

In one embodiment, compositions according to the subject invention canbe ingested by a subject in order for the compositions to be contactedwith the gastrointestinal tract (e.g., the target site) and have adesired local effect therein or to be absorbed therein for systemiceffects. Administration can also be achieved through, for example,injection (e.g., intravenous (IV), intramuscular (IM), intraperitoneal,intrathecal or subcutaneous), transdermal, rectal, urogenital (e.g.,vaginal), ocular, aural, nasal, mucosal, inhalation and cutaneousroutes.

In one embodiment, the health-promoting compound is a supplement. Thesupplement can be synthetic, or can be naturally-derived, for example,originating from microbial, fungal, plant or animal sources. Thesupplement can be a dietary and/or nutritional supplement, for example,providing nutrients such as vitamins (e.g., A (retinoids), B1(thiamine), B2 (riboflavin), B3 (niacin), B5 (panthothenic acid), B6(pyridoxine), B9 (folic acid), B12 (cobalamin), C (ascorbic acid), D(calciferol), E (tocopherol), H (biotin), K, and/or derivativesthereof); electrolytes and minerals (e.g., calcium, phosphorous,magnesium, potassium, sodium chloride, iodine, zinc, iron, copper,chromium, fluoride, selenium, manganese, and molybdenum); and fats,carbohydrates and/or proteins (e.g., whey, hemp, soy, collagen, aminoacids). The supplement can be a source of energy, alertness, and/orincreased physical performance, providing, for example, caffeine, yerbamate, creatine and/or guarana. The supplement can also be a botanical orherbal supplement, for example, turmeric root or ginseng, for holistichealth benefits.

In one embodiment, the health-promoting compound is water, wherein theadjuvant composition can be administered as an enhanced hydration orrehydration compound to increase the bioavailability and absorption ofwater in the GI tract.

In one embodiment, the health-promoting compound is a pharmaceutical orbiopharmaceutical. As used herein, the phrase “pharmaceutical” refers toa compound manufactured for use as a medicinal and/or therapeutic drug,whether prescribed by a health care professional or available over thecounter. As used herein, the phrase “biopharmaceutical” refers to abiological macromolecule or cellular component, such as a blood product,used as a pharmaceutical. Biopharmaceuticals are typically manufacturedin, extracted from, or semi-synthesized from biological sources.

In one embodiment, the pharmaceutical is selected from an antiviral(e.g., acyclovir or valacyclovir), an antibiotic (e.g. erythromycin),and a pain-reliever and/or anti-inflammatory compound (e.g., ibuprofenor aspirin).

Additional, and non-limiting examples of pharmaceuticals that can behealth-promoting compounds according to the subject invention include,analgesics (e.g., NSAIDs, opioids, acetaminophen, naproxen and localanesthetics); muscle relaxants; digestive aids (e.g., antacids, refluxsuppressants, PPIs, laxatives, probiotics, prebiotics, andantidiarrheals); cardiovascular drugs (e.g., beta blockers, calciumchannel blockers, diuretics, vasoconstrictors, vasodilators, cardiacglycosides, antiarrhythmics, nitrates); blood pressure/hypertensiondrugs (e.g., ACE inhibitors, alpha blockers, angiotensin receptorblockers); coagulation drugs (e.g., anticoagulants, heparin,antiplatelet drugs, fibrinolytics, anti-hemophilic factors andhaemostatic drugs); statins (e.g., LDL cholesterol inhibitors andhypolipidaemic agents); endocrine aids (e.g., androgens, antiandrogens,estrogens, gonadotropin, corticosteroids, HGH, vasopressin);antidiabetics (e.g., sulfonylureas, biguanides, metformin,thiazolidinediones, insulin); thyroid hormones and antithyroid drugs;

urogenital system drugs (e.g., antifungals, alkalinizing agents,quinolones, antibiotics, cholinergics, anticholinergics, fertilitymedications, hormonal contraceptives); central nervous system drugs(e.g., psychedelics, hypnotics, anesthetics, antipsychotics, eugeroics,antidepressants (including tricyclics, monoamine oxidase inhibitors,lithium salts, and SSRIs), antiemetics, anticonvulsants/antiepileptics,stimulants, amphetamines, dopamine agonists, antihistamines,cannabinoids, 5-HT antagonists); ocular medications (e.g., topicalanesthetics, sympathomimetics, parasympatholytics, mydriatics,cycloplegics, mast cell inhibitors); antimicrobials (e.g., antibiotics,antibacterials, antifungals, antiparasitics, antiprotozoals,amoebicides); antivirals (e.g., acyclovir, ribavirin, valacyclovir,famciclovir), antihistamines, anticholinergics, antiseptics,cerumenolytics, bronchodilators, antitussives, mucolytics,decongestants, antimalarials, antitoxins, antivenoms, vaccines,immunoglobulins, immunosuppressants, interferons, monoclonal antibodies,chemotherapeutic drugs and/or any other category of compounds that arecapable of treating any health condition, disease or disorder, or ofenhancing health in any way.

1. A composition with enhanced bioavailability of a health-promotingsubstance wherein said composition comprises a health-promotingsubstance selected from pharmaceuticals and nutritional supplements, andwherein said composition further comprises a bioavailability-enhancingamount of a biosurfactant adjuvant.
 2. The composition of claim 1,further comprising a pharmaceutically-acceptable carrier.
 3. Thecomposition of claim 1, wherein the biosurfactant is selected fromglycolipids and lipopeptides.
 4. The composition of claim 3, wherein theglycolipids are selected from sophorolipids, rhamnolipids, trehaloselipids, and mannosylerythritol lipids.
 5. The composition of claim 3,wherein the lipopeptides are selected from surfactins, iturins,lichenysins, and fengycin.
 6. The composition of claim 1, wherein thebiosurfactant is present in critical micelle concentration (CMC).
 7. Thecomposition of claim 1, formulated for administration to a subjectthrough a route selected from oral, intravenous, rectal, vaginal,ocular, transdermal, aural, nasal, mucosal, inhalation and cutaneous. 8.The composition of claim 1, formulated as a delivery system wherein thehealth-promoting substance is encapsulated in a nanocapsule formed bythe biosurfactant.
 9. A method for enhancing the bioavailability of ahealth-promoting substance, the method comprising administering to thesubject a therapeutically-effective amount of said substancesimultaneously or sequentially with an adjuvant composition comprising abiosurfactant.
 10. The method of claim 9, wherein the health-promotingsubstance and the adjuvant composition are administered by oralingestion, injection, or inhalation.
 11. The method of claim 9, used toenhance the penetration of the health-promoting substance intoepithelial cells in the subject's gastrointestinal tract.
 12. The methodof claim 9, used to enhance penetration of the health-promotingsubstance across the subject's blood-brain barrier.
 13. The method ofclaim 9, used to reduce the dose of the health-promoting substance thatis required for it to be therapeutically-effective in the subject.
 14. Amethod for enhancing the bioavailability of a health-promotingsubstance, the method comprising administering atherapeutically-effective amount of a composition of claim 1 to asubject in need thereof.
 15. The method of claim 14, used to enhance thepenetration of the health-promoting substance into epithelial cells inthe subject's gastrointestinal tract.
 16. The method of claim 14, usedto enhance penetration of the health-promoting substance across thesubject's blood-brain barrier.
 17. The method of claim 14, used toreduce the dose of the health-promoting substance that is required forit to be therapeutically-effective in the subject.